Methods for treating or preventing HBV infection or HBV related diseases

Abstract

The present invention relates to polymerase HBV mutant polypeptides comprising a mutated polymerase domain which is functionally disrupted for polymerase activity and fusion proteins comprising such polymerase mutant polypeptide. The present invention also relates to a nucleic acid molecule and an expression vector for expressing said polymerase mutant polypeptide as well as a composition which can be used for eliciting an immune response to HBV with the goal of providing a protective or therapeutic effect against HBV infection.

The invention claimed is: 
     
       1. A method for treating or inhibiting or delaying an HBV infection, a chronic HBV infection or HBV-associated liver lesions, liver cancer, liver inflammation, chronic liver disease, comprising one or more administration(s), to a subject in need thereof, of a therapeutically effective amount of composition comprising a nucleic acid molecule coding for a mutant polypeptide, a vector comprising said nucleic acid molecule, a host cell comprising said nucleic acid molecule or said vector, or any combination thereof,
 wherein said mutant polypeptide comprises a mutated HBV polymerase domain with an internal deletion that functionally disrupts the polymerase activity, wherein said internal deletion is of at least 4 amino acid residues and at most 30 amino acid residues, said mutated polymerase domain comprising:
 the amino acid sequence shown in SEQ ID NO:1 but lacking at least the Tyr residue in position 203, the Met residue in position 204, the Asp residue in position 205, the Asp residue in position 206, the Val residue in position 207, the Val residue in position 208, and the Leu residue in position 209. 
 
 
     
     
       2. The method according to  claim 1 , wherein said nucleic acid molecule encodes a mutant polypeptide which, in addition to the deletion of residues 203 to 209 of SEQ ID NO:1:
 comprises a polymerase domain having the amino acid sequence shown in SEQ ID NO:2; 
 comprises a RNaseH domain having the amino acid sequence shown in SEQ ID NO:3 or SEQ ID NO:4; 
 comprises an amino acid sequence which exhibits at least 90% of identity with the amino acid sequence shown in SEQ ID NO:5; or 
 comprises an amino acid sequence selected from which exhibits at least 90% of identity with the amino acid sequence shown in any of SEQ ID NOs 6-12. 
 
     
     
       3. The method according to  claim 1 , wherein said nucleic acid molecule encodes a mutant polypeptide which, in addition to the deletion of residues 203 to 209 of SEQ ID NO:1, comprises a nucleotide sequence exhibiting at least 90% of identity with the nucleotide sequence shown in any of SEQ ID NOs 13-17. 
     
     
       4. The method according to  claim 1 , wherein said vector is a plasmid or a viral vector for expression in higher eukaryotic cells or organisms. 
     
     
       5. The method according to  claim 4 , wherein said vector is a viral vector originating from a retrovirus, adenovirus, adenovirus-associated virus (AAV), poxvirus, herpes virus, measles virus, foamy virus, alphavirus, or vesicular stomatis virus. 
     
     
       6. The method according to  claim 5 , wherein said vector is a replication-defective adenoviral vector originating from a human or from a chimpanzee adenovirus. 
     
     
       7. The method according to  claim 6 , wherein the nucleic acid molecule is inserted in the adenoviral E1 region and placed under the control of a CMV promoter. 
     
     
       8. The method according to  claim 5 , wherein said vector is a poxviral vector originating from a canarypox, a fowlpox, or a vaccinia virus. 
     
     
       9. The method according to  claim 1 , wherein said vector is selected from the group consisting of:
 A defective Ad vector comprising inserted in place of the E1 region a nucleic acid molecule placed under the control of a promoter such as the CMV promoter, and encoding a mutant polypeptide comprising an amino acid sequence as shown in SEQ ID NO:5 or a fusion protein comprising an amino acid sequence as shown in SEQ ID NO:6 or SEQ ID NO:8; 
 A replication-defective Ad vector comprising inserted in place of the E1 region a nucleic acid molecule placed under the control of a promoter such as the CMV promoter, and comprising the nucleotide sequence shown in SEQ ID NO:13, SEQ ID NO:14 or SEQ ID NO:15; 
 A replication-defective Ad vector, especially a defective AdCh3 comprising inserted in place of the E1 region a nucleic acid molecule placed under the control of a promoter such as the CMV promoter and comprising the nucleotide sequence shown in SEQ ID NO:16 or SEQ ID NO:17; 
 A MVA vector comprising a nucleic acid molecule placed under the control of a vaccinia promoter such as the 7.5K or pH5R promoter, and encoding a mutant polypeptide comprising an amino acid sequence as shown in SEQ ID NO:5 or SEQ ID NO:10 or a fusion protein comprising an amino acid sequence as shown in SEQ ID NO:6, SEQ ID NO:8 or SEQ ID NO:12; and 
 A MVA vector comprising a nucleic acid molecule placed under the control of a vaccinia promoter such as the 7.5K or pH5R promoter, and comprising the nucleotide sequence shown in SEQ ID NO:13, SEQ ID NO:14 or SEQ ID NO:15. 
 
     
     
       10. The method according to  claim 1 , wherein said vector is in the form of infectious viral particles. 
     
     
       11. The method according to  claim 1 , wherein the mutant polypeptide encoded by said nucleic acid molecule is fused in frame to a signal peptide and to a trans-membrane peptide. 
     
     
       12. The method according to  claim 1 , wherein said composition further comprises a pharmaceutically acceptable vehicle. 
     
     
       13. The method according to  claim 1 , wherein said composition is formulated for intramuscular, subcutaneous, intradermal administration or scarification. 
     
     
       14. The method according to  claim 1 , wherein said composition comprises doses of about 5×10 8 , about 10 9 , about 5×10 9 , about 10 10 , about 5×10 10  vp or about 10 11  vp of an adenoviral vector. 
     
     
       15. The method according to  claim 1 , wherein said composition comprises doses of about 5×10 8 , about 10 7 , about 5×10 7 , about 10 8 , or about 5×10 8  pfu of an MVA vector. 
     
     
       16. The method according to  claim 1 , for treating a chronic HBV infection. 
     
     
       17. The method according to  claim 1 , for eliciting or stimulating an immune response in the treated organism. 
     
     
       18. The method according to  claim 17 , wherein said elicited or stimulated immune response is specific and/or non-specific, humoral and/or cellular. 
     
     
       19. The method according to  claim 18 , wherein said immune response is a T cell response CD4+ or CD8+-mediated or both, directed to an HBV polypeptide/epitope. 
     
     
       20. The method according to  claim 1 , wherein said vector is an adenoviral vector and said method comprises one or two intramuscular or subcutaneous administrations. 
     
     
       21. The method according to  claim 1 , which is carried out in combination with the standard of care. 
     
     
       22. The method according to  claim 1 , which is carried out according to prime boost modality. 
     
     
       23. The method according to  claim 22 , wherein the priming is carried out with an MVA vector and the boosting with an Ad vector. 
     
     
       24. The method according to  claim 23 , wherein the MVA and/or the Ad vector encodes the fusion protein shown in SEQ ID NO: 8. 
     
     
       25. The method according to  claim 24 , comprising at least 3 subcutaneous administrations of the MVA vector separated by a period of time varying from 3 days to 3 months followed by an intramuscular or subcutaneous boost of the adenovirus vector. 
     
     
       26. The method according to  claim 1 , wherein said HBV-associated liver lesions, liver cancer, liver inflammation, chronic liver disease is selected from the group consisting of: cirrhosis, steatosis, fibrosis, hepatocellular carcinoma and liver carcinoma.